Differential proteomics research of Bacillus amyloliquefaciens and its genome-shuffled saltant for improving fengycin production

Zhao, Junfeng; Zhang, Chong

doi:10.1016/j.bjm.2018.04.010

Cited by 7 publications

(4 citation statements)

References 20 publications

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“…Deletion of spo0E and rapA could both significantly increase the antifungal activity in WH1. Thereby, Spo0A is essential for biosynthesis of antifungal lipopeptides, and maintaining a certain level of Spo0A∼P is favorable for strengthening the antifungal activity in B. amyloliquefaciens , consistent with the previous reports that biosynthesis of fengycin and iturin is regulated by Spo0A in B. subtilis ( Rahman et al, 2006 ; Zhang et al, 2016 ; Zhao et al, 2018 ). On this basis, we deleted spo0E and rapA in Δ kinA Δ bdh Δ dhbF , respectively, but only deletion of rapA could further increase the antifungal activity in Δ kinA Δ bdh Δ dhbF .…”

Section: Discussionsupporting

confidence: 91%

Systemically engineering Bacillus amyloliquefaciens for increasing its antifungal activity and green antifungal lipopeptides production

Wang

Wang²,

Zhao

et al. 2022

Front. Bioeng. Biotechnol.

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The biosynthesis of antifungal lipopeptides iturin and fengycin has attracted broad interest; however, there is a bottleneck in its low yield in wild strains. Because the key metabolic mechanisms in the lipopeptides synthesis pathway remain unclear, genetic engineering approaches are all ending up with a single or a few gene modifications. The aim of this study is to develop a systematic engineering approach to improve the antifungal activity and biosynthesis of iturin and fengycin in Bacillus amyloliquefaciens. First, blocking the carbon overflow metabolic pathway to increase precursor supply of the branched-chain amino acids by knockout of bdh, disrupting sporulation to extend the stage for producing antifungal lipopeptides by deletion of kinA, blocking of siderophore synthesis to enhance the availability of amino acids and fatty acids by deletion of dhbF, and increasing Spo0A∼P by deletion of rapA, could improve the antifungal activity by 24%, 10%, 13% and 18%, respectively. Second, the double knockout strain ΔbdhΔkinA, triple knockout strain ΔbdhΔkinAΔdhbF and quadruple knockout strain ΔkinAΔbdhΔdhbFΔrapA could improve the antifungal activity by 38%, 44% and 53%, respectively. Finally, overexpression of sfp in ΔkinAΔbdhΔdhbFΔrapA further increased the antifungal activity by 65%. After purifying iturin and fengycin as standards for quantitative analysis of lipopeptides, we found the iturin titer was 17.0 mg/L in the final engineered strain, which was 3.2-fold of the original strain. After fermentation optimization, the titer of iturin and fengycin reached 31.1 mg/L and 175.3 mg/L in flask, and 123.5 mg/L and 1200.8 mg/L in bioreactor. Compared to the original strain, the iturin and fengycin titer in bioreactor increased by 22.8-fold and 15.9-fold in the final engineered strain, respectively. This study may pave the way for the commercial production of green antifungal lipopeptides, and is also favorable for understanding the regulatory and biosynthetic mechanism of iturin and fengycin.

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Section: Discussionsupporting

confidence: 91%

Systemically engineering Bacillus amyloliquefaciens for increasing its antifungal activity and green antifungal lipopeptides production

Wang

Wang²,

Zhao

et al. 2022

Front. Bioeng. Biotechnol.

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“…Moreover, some of the recent findings also show high potential in peptide synthesis, including the synthesis of some of the glucagon-like peptide by B. amyloliquefaciens strains, 7 synthesis of a bioactive peptide called fengycin for food applications, 82 and other peptides and proteins with different functions.…”

Section: Synthesis Of Essential Micro-and Macromoleculesmentioning

confidence: 99%

Prebiotic, Probiotic, Antimicrobial, and Functional Food Applications of Bacillus amyloliquefaciens

Yohannes

Wan

et al. 2020

J. Agric. Food Chem.

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Bacillus amyloliquefaciens belongs to the genus Bacillus and family Baciliaceae. It is ubiquitously found in food, plants, animals, soil, and in different environments. In this review, the application of B. amyloliquefaciens in probiotic and prebiotic microbes in fermentation, synthesis, and hydrolysis of food compounds is discussed as well as further insights into its potential application and gaps. B. amyloliquefaciens is also a potential microbe in the synthesis of bioactive compounds including peptides and exopolysaccharides. In addition, it can synthesize antimicrobial compounds (e.g., Fengycin, and Bacillomycin Lb), which makes its novelty in the food sector greater. Moreover, it imparts and improves the functional, sensory, and shelf life of the end products. The hydrolysis of complex compounds including insoluble proteins, carbohydrates, fibers, hemicellulose, and lignans also shows that B. amyloliquefaciens is a multifunctional and potential microbe which can be applied in the food industry and in functional food processing.

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“…Many studies have reported that genome shuffling and fermentation optimization are the two efficient approaches to improve lipopeptide production. For example, the fengycin yield of the Bacillus amyloliquefaciens mutant strain FMB72 was 8.30 times higher than that of the wild ES-2-4 strain after two rounds of genome shuffling (Zhao et al, 2016(Zhao et al, , 2018; the fengycin production of Bacillus subtilis F29-3 was increased from 1.2 to 3.5 g/L using the response surface methodology (RSM) optimization strategy (Wei et al, 2010). The addition of metal ions (Mn 2+ , Fe 2+ , Fe 3+ and Cu 2+ ) to culture media has been reported to significantly promote lipopeptide production in Bacillus strains (Zhao et al, 2013).…”

Section: Introductionmentioning

confidence: 99%

Effect of metal ions on lipopeptide secretion from Bacillus subtilis strain FJAT-4: Negative regulation by Ca2+

Chen

Zheng

Chen

et al. 2022

Journal of Applied Microbiology

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Aims: This study aimed to investigate the effect of metal ions on lipopeptide production by Bacillus subtilis strain FJAT-4 and the mechanism of negative regulation by Ca 2+ . Methods and Results:The quantitative measurement of lipopeptides in response to K + , Na + , Mg 2+ and Ca 2+ addition was carried out by LC-MS. The contents of fengycin and surfactin varied within the range of 116. 24-129.80 mg/L and 34.03-63.11 mg/L in the culture media containing K + , Na + and Mg 2+ , while the levels were 0.86 and 0.63 mg/L in the media containing Ca 2+ . Ca 2+ at a high concentration (45 mM) did not adversely affect the growth of strain FJAT-4, but caused significant downregulation of lipopeptide synthesis-related gene expression, corresponding to a decrease in lipopeptide production. This inhibition by Ca 2+ was further investigated by proteomic analysis. In total, 112 proteins were upregulated and 524 proteins were downregulated in the presence of additional Ca 2+ (45 mM). Among these differentially expressed proteins (DEPs), 28 were related to phosphotransferase activity, and 42 were related to kinase activity. The proteomics results suggested that altered levels of three two-component signal-transduction systems (ResD/ResE, PhoP/PhoR and DegU/DegS) might be involved in the control of expression of the fen and srfA operons of FJAT-4 under high calcium stress. Conclusions:The Ca 2+ at the high concentration (45 mM) triggers a decrease in lipopeptide production, which might be attributed to the regulation of three twocomponent signal-transduction systems ResD/ResE, PhoP/PhoR and DegU/DegS. Significance and Impact of the Study:The regulatory effect of calcium on the expression of genes encoding lipopeptide synthetases can be applied to optimize the production of lipopeptides.

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Differential proteomics research of Bacillus amyloliquefaciens and its genome-shuffled saltant for improving fengycin production

Cited by 7 publications

References 20 publications

Systemically engineering Bacillus amyloliquefaciens for increasing its antifungal activity and green antifungal lipopeptides production

Systemically engineering Bacillus amyloliquefaciens for increasing its antifungal activity and green antifungal lipopeptides production

Prebiotic, Probiotic, Antimicrobial, and Functional Food Applications of Bacillus amyloliquefaciens

Effect of metal ions on lipopeptide secretion from Bacillus subtilis strain FJAT-4: Negative regulation by Ca2+

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